antigen-antibody reactions. antigen-antibody interactions: are reversible specific non-covalent...
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Antigen-antibody interactions:Are reversible specific non-covalent biochemical reactions:– Hydrogen bonds (A chemical bond in which a hydrogen atom of one
molecule is attracted to an electronegative atom of another molecule)– Electrostatic bonds(A valence bond in which two atoms, attracted by
electrostatic forces, transfer one or more electrons between atoms)– Van der Waal forces (forces acting between nonbonded atoms or
molecules)– Hydrophobic bonds(The attractive force between molecules due to the
close positioning of non-hydrophilic portions of the two molecules
Can be represented by the formula:
K1=constant of association K2=constant of dissociation
Ag + Ab Ag Ab K1
K2
The affinity: is the strength of the reaction between a single antigenic
determinant and a single combining site on the antibodyor it is the association constant for binding (KA)
KA= k1/k2
Valence: the number of epitopesAvidity: is the collective affinity of multiple binding
sites(affinity+ Valence)
Types of Antigen-antibody reactions :
–Precipitation
–Agglutination
–Neutralization (Antitoxins)
–Opsonization–Antibody-dependant cell-mediated
cytotoxicity–The complement activation Membrane
attack complex
PRECIPITATION Is the reaction of soluble Ag with soluble Ab.The reaction results in the formation of Ag-Ab
complexes (lattices)
AntigenAntigen
AntibodAntibodyy
The Quantitative Precipitation Reaction:Varying amounts of Ag are mixed and incubated with
Constant volume of antiseraPrecipitate is measured, amount of precipitate depends
on : o the ratio of Ag : Abo The Ab avidity
Plot in a curve, three zones are detected:i. Zone of Ag excess : insufficient Ab too small complexes to
precipitate ii. Equivalence zone : large lattice is formed visible
precipitatesiii. Zone of Ab excess : not enough Ag too small complexes to
precipitate
AGGLUTINATIONAbs can bind and cross-link cells or particles
aggregate formationEntrap microbial invadersIgM & IgA are the most suitable (IgG in sufficient amounts
can agglutinate cells)
Agglutination
RBC
RBC
RBC
RBC
IgM AntibodyIgM Antibody
IgG AntibodyIgG Antibody
RBC
RBC RBC
RBC RBC
RBC RBC
Applications of Agglutination 1. Agglutination/Hemagglutination: a. Qualitative agglutination test Determination of blood types or antibodies to blood group Ags b. Quantitative agglutination testAgglutination tests can also be used to measure the level of
antibodies to particulate antigens.(titration)2. Passive hemagglutination: erythrocytes are coated
with a soluble antigen (e.g. viral antigen, a polysaccharide or a hapten) and use the coated red blood cells in an agglutination test for antibody to the soluble antigen
3. Coomb's Test (Antiglobulin Test)
NEUTRALIZATIONIs the binding of Ab to microbial epitopes or soluble
molecules(e.g. toxins) which inhibits their binding to host cells.
Abs are mostly IgG & IgAUsed to identify toxins and viruses
OPSONIZATIONIs the process by which a pathogen is marked (tagged)
for ingestion and destruction by phagocytic cells
Antibody-dependant cell-mediated cytotoxicity• Coating of an organism can attract phagocytic cells
as well as other cytolytic cells(NK cells, eosinophils)
• The organism may be: bacteria, protozoa, parasitic worms
• These cells use cytolytic mechanisms to kill those organisms